A complete separation of lipids by three‐directional thin layer chromatography

Abstract

Abstract A novel three‐directional thin layer chromatography (TLC) method is reported by which all the polar and neutral lipids are isolated on a single TLC plate. Following resolution of the phospholipids by two‐directional TLC, lipids are visualized by ultraviolet light after spraying with 2′,7′‐dichloro‐fluorescein. A line is drawn across the plate, parallel to the second direction of development, separating the resolved phospholipids and the neutral lipids concentrated along the solvent front. The TLC plate is then chromatographed in the reverse direction of the second development to resolve the neutral lipids. By exposing the lipids to HCl fumes after the first development, the plasmalogen content of the lipids may also be determined. This new technique is rapid and lends itself to qualitative and quantitative analyses of total lipids.